The study results of in vitro formation of tissue-engineered cartilage construct on the basis of cell-engineered construct composed of biopolymer hydrogel matrix and human adipose tissue-derived mesenchymal stromal cells (hADSCs) are presented. It was revealed that hADSCs in biopolymer hydrogel matrix Sphero®GEL under chondrogenic conditions generate three-dimensional structures and produce cartilaginous extracellular matrix components: collagen type II and glycosaminoglycans.
Adhesion and proliferation of NIH/3Т3 mouse fibroblasts on the surfaces of bacterial copolymer poly(3-hydroxybutyrate-co-3-hydroxyvalerate) and its mixture with poly(ethylene glycol) with differing crystallinity, surface energy, and mean roughness was investigated. The surface mean roughness of all films on both sides, at the air interface and at the glass interface, measured by atomic force microscopy, was higher (from 17.0±1.4 nm to 290.8±7.0 nm) than that of the tissue culture polystyrene control (9.5±0.6 nm). The structure, surface energy, and chemical composition of bacterial films were studied by differential scanning calorimetry, contact angle measurements, and Fourier transform infrared (FTIR) spectroscopy. After 48 h, cell proliferation on all surfaces was significantly less than on the control substrate; however, after 72 and 96 h, cell proliferation was comparable with control on some surfaces with sufficient roughness. Addition of poly(ethylene glycol) resulted in an increase of adhesion and the metabolic activity of the cells, even for relatively smooth surfaces. The complex correlation of cell metabolic activity with surface energy and crystallinity for "rough" (mean roughness >100 nm) and "smooth" (mean roughness < 100 nm) surfaces is discussed.
Objective: to develop a method for modifying composite small-diameter porous tubular biopolymer scaffolds based on bacterial copolymer poly(3-hydroxybutyrate-co-3-hydroxyvalerate) and gelatin modified with a double-layered bioactive coating based on heparin (Hp) and platelet lysate (PL) that promote adhesion and proliferation of cell cultures.Materials and methods. Composite porous tubular biopolymer scaffolds with 4 mm internal diameter were made by electrospinning from a 1 : 2 (by volume) mixture of a 10% solution of poly(3-hydroxybutyrateco- 3-hydroxyvalerate) copolymer, commonly known as PHBV, and a 10% solution of gelatin, respectively, in hexafluoro-2-propanol. The structure of the scaffolds was stabilized with glutaraldehyde vapor. The scaffolds were modified with a bioactive Hp + PL-based coating. The surface morphology of the samples was analyzed using scanning electron microscopy. Biological safety of the modified scaffolds in vitro (hemolysis, cytotoxicity) was evaluated based on the GOST ISO 10993 standard. Interaction with cultures of human endothelial cell line (EA. hy926) and human adipose-derived mesenchymal stem cells (hADMSCs) was studied using vital dyes.Results. We developed a method for modifying small-diameter composite porous tubular biopolymer scaffolds obtained by electrospinning from a mixture of PHBV and gelatin modified with double-layered bioactive coating based on covalently immobilized Hp and human PL. The modified scaffold was shown to have no cytotoxicity and hemolytic activity in vitro. It was also demonstrated that the developed coating promotes hADMSC adhesion and proliferation on the external surface and EA.hy926 on the internal surface of the composite porous tubular biopolymer scaffolds in vitro.Conclusion. The developed coating can be used for the formation of in vivo tissueengineered small-diameter vascular grafts.
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